ETS2-dependent control in cardiomyocyte ischemia/reperfusion injury

项目总结/摘要 心力衰竭是世界范围内发病率和死亡率的主要原因，并且是心血管疾病的主要原因。 射血分数降低的心力衰竭（HFrEF）是冠状动脉疾病。丝裂原活化 蛋白激酶（MAPK）是一种重要的信号转导级联反应，在两种细胞 生长和细胞死亡。MAPK通路的MEK 1-ERK 1/2分支已显示促进 在心脏中的生理和病理生长，以及防止凋亡细胞死亡后， 缺血/再灌注（I/R）损伤。我们以前已经证明，转录因子ETS 2，a E26转化特异性序列（ETS）-结构域家族的成员，被磷酸化并被激活， Erk 1/2在肥大刺激后。我们的初步数据显示心肌细胞- 在I/R和永久性结扎中，ETS 2的特异性丢失导致对缺血性损伤的易感性增加 （心肌梗塞）心力衰竭模型。连接蛋白43（Cx43），主要的间隙连接通道- 在心肌细胞中形成蛋白，已经被认为在缺血性损伤和缺血性 预处理我们的初步数据表明，ETS 2激活Cx43转录，Cx43是 在缺乏ETS 2的情况下下调。我们将检验ERK 1/2/ETS 2通路保护细胞凋亡的假设。 部分通过上调Cx43对抗I/R损伤。 目的1：探讨ERK 1/2-ETS 2通路在缺血再灌注损伤中的作用。我们的初步数据显示 ETS 2保护I/R损伤的模型。我们将确认并延长这一点，使用损失和收益的- 功能的方法在体内和体外。我们将跟踪ERK激活ETS 2的时间，以及ERK对ETS 2的反应。 每一个急性和长期重塑。 目的2：探讨ETS 2对I/R损伤中Cx43表达和功能的影响。我们的数据 提示Cx43在缺血性损伤中起保护作用。我们的初步数据还表明，ETS 2是 Cx43基因表达的直接转录调节因子。我们将确认并扩展这些发现， 功能丧失和功能获得两种方法。我们还将确定ETS 2在Cx43介导的细胞凋亡中的作用。 IPC中的心脏保护。 目的3：确定ETS 2在心肌缺血条件下的下游靶点和相互作用物。 应力我们将使用RNAseq和ChIPseq分析全基因组心脏基因表达， ETS 2下游基因靶向急性和长期I/R损伤。我们将使用免疫沉淀， 质谱技术来揭示新的蛋白质相互作用。

Project Summary/Abstract Heart failure is a leading cause of morbidity and mortality around the world, and the leading cause of heart failure with reduced ejection fraction (HFrEF) is coronary artery disease. The mitogen-activated protein kinases (MAPK) are an essential signal transduction cascade that play a central role in both cell growth and cell death. The MEK1-ERK1/2 branch of the MAPK pathway has been shown to promote both physiologic and pathologic growth in the heart, as well as protect against apoptotic cell death after ischemia/reperfusion (I/R) injury. We have previously demonstrated that the transcription factor ETS2, a member of E26 transformation-specific sequence (ETS)-domain family, is phosphorylated and activated by Erk1/2 upon hypertrophic stimulation. Going forward, our preliminary data reveal that cardiomyocyte- specific loss of ETS2 results in increased susceptibility to ischemic injury in both I/R and permanent ligation (myocardial infarction) models of heart failure. Connexin43 (Cx43), the predominant gap junction channel- forming protein in cardiomyocytes, has been suggested to play a role in both ischemic damage and ischemic preconditioning. Our preliminary data show that ETS2 activates Cx43 transcription and that Cx43 is downregulated in the absence of ETS2. We will test the hypothesis that the ERK1/2/ETS2 pathway protects against I/R injury in part through the upregulation of Cx43. Aim 1: To determine the role of the ERK1/2-ETS2 pathway in I/R injury. Our preliminary data suggest a model in which ETS2 protects against I/R injury. We will confirm and extend this using loss- and gain-of- function approaches in vivo and in vitro. We will track the timing of ETS2 activation by ERK and the response of each acutely and in long-term remodeling. Aim 2: To determine the impact of ETS2 on Cx43 expression and function in I/R injury. Our data suggest that Cx43 plays a protective role in ischemic injury. Our preliminary data also suggest that ETS2 is a direct transcriptional regulator of Cx43 gene expression. We will confirm and extend these findings using both loss- and gain-of-function approaches. We will also determine the role of ETS2 in Cx43-mediated cardioprotection in IPC. Aim 3: To determine the downstream targets and interactors of ETS2 under conditions of cardiac stress. We will profile genome-wide cardiac gene expression using RNAseq and ChIPseq to determine ETS2 downstream gene targets in both acute and long-term I/R injury. We will use immunoprecipitation and mass spectrometry to unveil novel protein interactions.